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A new glycosyltransferase enzyme from family 91, ugt91p3, is responsible for the final glucosylation step of crocins in saffron (Crocus sativus l.)

TitleA new glycosyltransferase enzyme from family 91, ugt91p3, is responsible for the final glucosylation step of crocins in saffron (Crocus sativus l.)
Publication TypeArticolo su Rivista peer-reviewed
Year of Publication2021
AuthorsLópez, A.J., Frusciante Sarah, Niza E., Ahrazem O., Rubio-Moraga A., Diretto Gianfranco, and Gomez-Gomez L.
JournalInternational Journal of Molecular Sciences
Volume22
ISSN16616596
Keywordsbiosynthesis, Biosynthetic Pathways, Carotenoid, Carotenoids, chemistry, Computer simulation, crocin, Crocus, enzymology, gene expression profiling, gene expression regulation, genetics, Glycosylation, glycosyltransferase, Glycosyltransferases, metabolism, Plant, plant protein, Plant Proteins, procedures, two hybrid system, Two-Hybrid System Techniques
Abstract

Crocetin is an apocarotenoid formed from the oxidative cleavage of zeaxanthin, by the carotenoid cleavage enzymes CCD2 (in Crocus species) and specific CCD4 enzymes in Buddleja davidii and Gardenia jasminoides. Crocetin accumulates in the stigma of saffron in the form of glucosides and crocins, which contain one to five glucose molecules. Crocetin glycosylation was hypothesized to involve at least two enzymes from superfamily 1 UDP-sugar dependent glycosyl-transferases. One of them, UGT74AD1, produces crocins with one and two glucose molecules, which are substrates for a second UGT, which could belong to the UGT79, 91, or 94 families. An in silico search of Crocus transcriptomes revealed six candidate UGT genes from family 91. The transcript profiles of one of them, UGT91P3, matched the metabolite profile of crocin accumulation, and were co-expressed with UGT74AD1. In addition, both UGTs interact in a two-hybrid assay. Recombinant UGT91P3 produced mostly crocins with four and five glucose molecules in vitro, and in a combined transient expression assay with CCD2 and UGT74AD1 enzymes in Nicotiana benthamiana. These results suggest a role of UGT91P3 in the biosynthesis of highly glucosylated crocins in saffron, and that it represents the last missing gene in crocins biosynthesis. © 2021 by the authors. Licensee MDPI, Basel, Switzerland.

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URLhttps://www.scopus.com/inward/record.uri?eid=2-s2.0-85112512605&doi=10.3390%2fijms22168815&partnerID=40&md5=89805bd6207689f39532998632bcffc6
DOI10.3390/ijms22168815
Citation KeyLópez2021